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Grapevine leafroll disease (GLD) is an economically important virus disease affecting grapes (Vitis spp.) in almost all grape-producing areas, but little is known about its effect on metabolic constitutes of affected grapes. Fifty-seven suspicious grapevine samples of various cultivars showing symptoms of leaf curl, vein greening, reduced leaf size, necrosis, mosaics or reddening and reduced fruit yield and quality were collected during the summer of 2016–2017 in two regions of Azerbaijan and tested serologically (rapid one-step assay AgriStrip, DAS-ELISA) and molecularly (RT-PCR) for the presence of the major viruses infecting these crop. According to the result of serological tests, two cultivars (Saperavi and Narchicheyi) were found to be infected by GLRaV-3. RT-PCR using specific primers (LR3-8504V and LR3-9445C) for a part of the GLRaV-3 viral coat protein gene confirmed the presence of the virus by amplification of expected 942 bp fragments. The levels of some metabolic constituents were studied comparatively in the leaves of GLRaV3-infected and non-infected grapes and an increase of tocopherols, soluble sugars, total phenols, soluble protein, proline and glycine betaine was detected in infected grapes. Thus, it can be considered as an adaptive ability of various grapevine varieties against pathogen infection.